Combining resources to obtain a comprehensive survey of the bovine embryo transcriptome through deep sequencing and microarrays

摘要: While most assisted reproductive technologies (ART) are considered routine for the reproduction of species economical importance, such as bovine, impact these manipulations on developing embryo remains largely unknown. In an effort to obtain a comprehensive survey bovine transcriptome and how it is modified by ART, resources were combined design embryo-specific microarray. Close one million high-quality reads produced from subtracted libraries using Roche 454 Titanium deep sequencing technology, which enabled creation augmented genome catalog. This catalog was enriched with transcripts, included newly discovered indel type 3'UTR variants. Using this catalog, EmbryoGENE Bovine Microarray designed composed total 42,242 probes, including 21,139 known reference genes; 9,322 probes novel transcribed regions (NTRs); 3,677 alternatively spliced exons; 3,353 3'-tiling probes; 3,723 controls. A suite bioinformatics tools also developed facilitate microrarray data analysis database creation; includes quality control module, Laboratory Information Management System (LIMS) microarray software. Results obtained during study have already led identification differentially expressed blastocyst targets, NTRs, splice variants type, We able confirm results real-time PCR, indicating that has power detect physiologically relevant changes in gene expression.